A quartz crystal microbalance sensor coated with MIP for

Bisphenol A and its properties

Naoto Tsuru
*
, Masashi Kikuchi, Haruma Kawaguchi
c
, Seimei Shiratori
a,b
a
Department of Applied Physics & Physico-informatics, Faculty of Science & Technology, Keio University, 3-14-1 Hiyoshi,
Kohoku-ku, Yokohama-shi 223-8522, Japan
b
SNT Co. Ltd., 308-10, Ogura, Saiwai-ku, Kawasaki-shi, 212-0054, Japan
c
Department of Chemistry, Faculty of Science and Technology, Keio University 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama-city,
Kanagawa-pref., 223-8522, Japan
Abstract
Molecular imprinted polymer coated quartz crystal microbalance (MIP-QCM) sensors were fabricated for the detection of bisphenol A
in the liquid phase. The MIP-QCM sensor was designed for the selective detection of BPA, and MIP was designed for the proper adhesion
between MIP and QCM Au electrode. MIP, contained specific cavities by self organized structure of functional monomers around a
template, was synthesized on Au electrode coated with the self-assembled monolayer (SAM) of QCM. 2-Aminoethanethiol (AET) and 11-
mercaptoundecanoic acid (MUA) were used as SAM. SAM had ion-pair interactions with functional monomers to form proper adhesions
between MIP and QCM electrode. The MIP-QCM sensor performances such as sensitivity and selectivity are largely influenced by types of
materials and compositions, amount of MIP, solvent, and SAM. Through investigations of these influences, sensor characteristics such as
sensitivity and selectivity are improved. These achievements of design were proved to be applicable for the fabrication of various MIP-
QCM sensors.
D 2005 Published by Elsevier B.V.
Keywords: MIP; QCM; BPA; Chemical sensor
0040-6090/$ - see front matter D 2005 Published by Elsevier B.V.
doi:10.1016/j.tsf.2005.07.005
1. Introduction
Bisphenol A (BPA) [4,4V -(1-methylethylidene) bisphenol]
shows weak estrogen mimic properties [1] and is recognized
as reproductive and as a developmental toxicity [2], never-
theless it is one of the most important intermediates in
industrially manufactured polycarbonates, epoxy and poly-
ester resins. BPA can be found in these resultant products
such as electrical and electronic equipment, food packing,
optical applications, films, motor parts and medical appli-
ances. These products are used in the imminent environment,
which means that BPA exist in our everyday lives. Almost all
the small amount of environmental BPA exposure can be
detected in industrial effluent, especially nearby paper
recycling factories, sewage-treatment plants and garbage
dumps [2]. Based on the idea of Pollutant Release and
Transfer Register (PRTR), major countries are trying to
control such environmental exposures. The European Union
has assessed a tolerance daily intake (TDI) of BPA at 0.01
mg/kg/day, as Japan assessed its TDI of BPA at 0.05 mg/kg/
day. Under Japanese food hygiene law, elution of BPA to
food from dish, which is manufactured by polycarbonate
resin, was regulated to be under 2.5 ppm. Hence, the
detection of BPA is very important in maintaining an
awareness of pollutants in our immediate environments.
Recently, various analytical methods have been
employed for the determination of BPA. Most of these
methods are based on high-performance liquid chromatog-
raphy (HPLC) [3] and Gas Chromatography/Mass Spec-
trometer (GC/MS) [4]. Enzyme-linked immunosorbent
assay (ELISA) has been useful as well [5]. These methods
are expensive, and require pretreatment. These have little
facility in measurement on site without pretreatment.
The research for highly selective, low-cost, stable,
sensitive, durable, and facile chemical sensors is an
attractive field. Quartz crystal microbalances are well * Corresponding author.
Thin Solid Films 499 (2006) 380 385
www.elsevier.com/locate/tsf
applicable to sensitive and selective detection, and portable
in situ measurement [6]. Compared to semiconductor gas
sensor, QCM sensors are considered to rather selective [7].
In this work, MIP was employed for QCM as selective
polymer membrane.
MIPs consist of functional monomers, cross-linkers and a
template. They have remarkable selectivity for template
because of their specific structure and characteristics. It is
formed by functional monomers self-organized around a
template by noncovalent interactions or reversible covalent
interactions during polymerization and coagulated with
cross-linkers. After polymerization, an extraction of a
template creates specific cavities by self-organized structure
of functional monomers around a template. Therefore, MIPs
possess excellent selective adsorption for templates due to
the stereoscopic shape of specific cavity and interactions. In
addition to this property, they are reusable and durable
against chemical conditions.
Such MIP properties are applicable for various fields. In
the field of HPLC study, MIPs have been successfully
developed in stationary phases in chromatography [8], and
solid-phase extraction [9]. Especially, the application of MIP
on QCM sensors is a promising field for new detection
systems.
MIP-QCM sensor technology has been progressing in the
past decade. Various methods are proposed to achieve MIP-
QCM sensors. The simplest suggested method is the direct
polymerization of MIP on the QCM electrode. Although it
was a simple procedure, this method proved that proper
adhesion could not be made between MIP and the QCM
electrode. And, another simple method was suggested where
a microscope coverslip was put onto the polymerization
solution [10]. This simple method was also reported that a
certain cross-linker concentrations led to an elimination of
MIP on the QCM electrode [11,12].
In our research, QCM Au electrode was coated with
SAM that had ion-pair interactions with functional mono-
mers of MIP. AET was used as SAM that had ion-pair
interactions with methacrylic acid, and MUAwas used with
4-vinyl pyridine. This method succeeded in the adhesion,
and achieved MIP-QCM sensor for BPA. Following topics
were discussed and designed for improvement of MIP-QCM
sensor beside the adhesion: Material types, its composition,
amount of MIP and presence of solvent and SAM. Through
these investigations, MIP-QCM sensor characteristics were
improved. Achievement of this study proved the certain
properties of MIP-QCM sensors, and gave guidance for
practical application of MIP-QCM sensors.
2. Experimental section
2.1. Materials
The cross-linkers ethylene glycol dimethacrylate (EGDMA),
the functional monomers methacrylic acid (MAA), the
initiator 2,2V -azobisisobutyronitrile (AIBN), ethanol, acetic
acid, phenol, 4,4V -diaminopheyhylmethane, aniline, metha-
nol, the SAM cysteamine Hydrochloride (AET) [2-Amino-
ethanethiol Hydrochloride], chloroform and functional
monomers 4-vinyl pyridine (4-Vpy) were purchased from
Wako Pure Chemicals. The SAM 11-mercaptoundecanoic
acid (MUA) and the template bisphenol A (BPA) were
purchased form Aldrich. All commercial chemicals were
used without further purification. Deionized water (18MM&
cm) was used as pure water.
2.2. Appartus
10 MHz AT-cut quartz crystals coated with Au were
used. One side of the Au electrode was glued by silicone
adhesive prior to use.
In this research, these crystal resonators fixed to an
oscillator circuit were dipped into stirred pure water 100 ml.
Frequency changes due to adding sample solution to stirred
water were continuously recorded by Lab-View software.
2.3. Preparation of MIP-QCM sensors
Before coating, the Au electrode was washed with 1.2 M
NaOH and 1.2 M HCl, respectively for 5 min, and then with
pure water. Next AET and MUA were freshly prepared as 1
mM solutions in absolute ethanol. The Au electrode was
fully immersed in the AET or MUA solution for 20 h, and
was washed by sonication 5 min with fresh ethanol solution.
Then QCM Au electrode coated with SAM. BPA as a
template was mixed with the functional monomer, the cross-
linker EGDMA and the solvent chloroform at ratios listed in
Table 1. The polymerization solution was deoxygenated
with nitrogen gas for 10 min, added with the AIBN initiator,
and the solution was cast directly on the QCM Au electrode
coated by SAM. Then the QCM was immediately placed in
a 60 -C oil bath for 6 h, while nitrogen purged. Fig. 1 shows
the preparation procedure of MIP on QCM gold electrode.
The resultant MIP on QCM was soaked for 6 h by a
methanol-acetic acid (15: 0.5 wt.%) solution to extract
template molecular BPA. Non imprinted polymer (NIP)
was fabricated by the composition 4 indicated in Table 1
without using BPA. Non imprinted polymer coated quartz
crystal microbalance (NIP-QCM) sensor was fabricated by
the above procedure excluding BPA.
Table 1
Polymerization solution variation and ratio
Number SAM Functional
monomer
Chloroform
(ml)
Ratio [mmol] (BPA: Functional
monomer: EGDMA)
1 AET MAA 0 1:4:20
2 Nothing MAA 0 1:4:20
3 AET MAA 10 1:4:20
4 MUA 4-Vpy 0 1:6:6
5 Nothing 4-Vpy 0 1:6:6
6 MUA 4-Vpy 10 1:6:6
N. Tsuru et al. / Thin Solid Films 499 (2006) 380385 381
2.4. Measurement
1 ml BPA water solutions of several concentrations were
prepared as sample solutions. The QCM adsorption meas-
urement was measured by frequency shifts due to adding
sample solutions to 100 ml of stirred pure water. These raw
frequency shifts were compensated by the frequency shifts
of 1 ml pure water.
3. Results and discussion
3.1. Formation of MIP
By comparing the results of composition 1 (Table 1)
and composition 2, effects of SAM were discussed. The
MIP-QCM sensor manufactured by composition 2 was
fabricated several times, but they hardly qualified as
sensor. The elimination of MIP from QCM electrode was
observed for some of them. Even if they had performed as
sensor, they showed little frequency shifts because they did
not have enough weight of membrane (Fig. 2). Meanwhile,
the MIP-QCM sensor manufactured by composition 1
acted as sensors, which proved that proper adhesion was
formed between functional monomers of MIP and SAM.
Therefore, the MIP-QCM sensor fabricated by the compo-
sition 2 shown in Table 1 did not have any relation
between MIP quantity and weight of membrane, but
composition 1 had almost a linear relation. These similar
phenomena also happened between composition 4 and 5.
The different sensor performance was observed. The MIP-
QCM sensor manufactured by composition 5 showed
greater sensor response than MIP-QCM sensor by compo-
sition 2. This was because the MIP consisted of 4-Vpy as
functional monomers having greater capability to catch
BPA than the MIP consisting of MAA. We concluded that
sensor performance was largely influenced by composition
of membrane. Even if both MIP-QCM sensors had very
small film amount, one performed as a sensor, and the
other did not.
When the results of composition 1 (Tables 1) were
compared to that of composition 3, effects of solvent could
be discussed. In the previous study, the type of solvent had
hardly influence on MIP properties [13]. Then we focused
on the necessity of solvent. First, we evaluated the MIP that
was synthesized without the solvent. The adsorption and
desorption property of MIP itself was observed by FT-IR,
and it was proved that the MIP without the solvent also had
adsorption and desorption property for target molecules.
Then two types of MIP composition 1 and 3 synthesized on
QCM electrode, and sensor responses were compared. The
MIP-QCM sensor manufactured by composition 3 was
fabricated several times, but it did not work as sensors at
times. On the contrary, the reproducibility of MIP-QCM
sensor manufactured by composition 1 was confirmed over
5 times. Consequently, we found that sensor fabricated by
composition 1 in the Table 1 showed better frequency shifts
than composition 3 that contains chloroform in this work.
NH
2
NH
2
NH
2
NH
2
NH
2
NH
2
NH
2
NH
2
Au electrode (1) (2)
BPA
COOH
COOH
C O O H
C
O
O
H C
O
O
H
Fig. 1. Preparation procedure of MIP on QCM gold electrode: (1) shows
that gold electrode was coated by SAM, and (2) shows the appearance
during polymerization.
Fig. 2. Frequency shifts vs. time. MIP-QCM sensor is curve 1, and NIP-QCM sensor is curve 2. Both weight of membrane is almost the same. BPA
concentration was 600 ppb.
N. Tsuru et al. / Thin Solid Films 499 (2006) 380385 382
These similar phenomena were also observed between
composition 4 and 6.
To discuss the location of the adsorption phenomenon,
we investigated the influence on MIP quantities. We
prepared several MIP-QCM sensors that had different
quantities of MIP, and found the different characteristics
in the frequency shifts. Fig. 3 shows that increasing the
weight of MIP led to improvement of MIP-QCM sensor
sensitivity until 1.5 Al casting quantity (approximately over
7000 Hz). This result proved that adsorption of MIP-QCM
sensors was carried out not only on the surface but also
inside the MIP film.
This MIP-QCM sensor reproducibility was confirmed
over 30 times.
3.2. Sensitivity
The most sensitive MIP-QCM sensor was manufactured
by composition 4, and its MIP quantity was 6430 Hz.
Detection limit was 100 ppb. The frequency shift curve of
this sensor is shown in Fig. 3. This MIP-QCM sensor took
110 min to respond 70 Hz at 600 ppb. The calibration
curve is shown in Fig. 4. Calibration curve are based on the
steady state frequency shifts after adding sample solution.
The reason of steep increase is not clarified yet, however
we considered that the physical adsorption started to the
surface of BPA which had already absorbed on the MIP
polymer.
3.3. Selectivity
Fig. 5 shows the selective property of MIP-QCM sensor
manufactured by composition 1 and 4. According to Fig. 5,
MIP-QCM sensor by composition 4 indicated better
selectivity than composition 1. Compared with frequency
shifts for analyte, composition 4 showed better discrim-
ination between BPA and others. We considered that this
result indicates that composition 4 has better selectivity than
Fig. 3. Frequency shifts for BPA of MIP-QCM sensor manufactured by composition 4 at each amount of membrane.
Fig. 4. Calibration curve: Consentrations(ppb) vs. Frequency shifts.
N. Tsuru et al. / Thin Solid Films 499 (2006) 380385 383
composition 1. Both tended to respond, depending on
analyte shapes. The more these analyte resembled BPA, the
bigger the frequency shifts. Then, Phenol most resembled
analyte with BPA in the point of shape and functional
moiety, therefore phenol showed the best frequency shifts
excluding BPA. These consequences proved that this
recognition was attributed to the specific shape and non-
covalent interactions. Then, it was considered that the
response of phenol was attributed to interaction, and the
response of 4,4-diaminodiphenylmethane to the shape.
Since the aniline did not have similar shapes with BPA
and there was no interaction with BPA. As shown is the
figure, aniline did not show any response to BPA. Therefore,
Fig. 5 also shows the effect of functional monomer. MIP-
QCM sensor by composition 4 showed better selectivity. It
is considered that this was originated by the difference of the
interaction. Composition 1 only had hydrogen bonding
interaction with BPA, however composition 4 had inter-
actions and hydrogen bonding interactions. Therefore,
composition 4 had stronger interactions with BPA, and it
was considered that stronger interaction originated better
imprinting cavity.
4. Conclusion
In conclusion, MIP was successfully coated on the QCM
Au electrode, because the adhesion between MIP and QCM
Au electrode well formed by SAM that had ion-pair
interactions with functional monomers in MIP. Then we
have achieved MIP-QCM sensor for BPA, and shown that
the MIP-QCM sensor can detect BPA selectively in the
liquid phase with a sensitivity of 100 ppb.
Through the fabricating design for MIP-QCM sensor, we
found that MIP-QCM sensor manufactured by composition
4 showed the best sensor characteristics. Our investigation
proved that (a) polymerization solution without solvent
showed better performance as sensor for this MIP-QCM
sensor procedure, (b) the QCM Au electrode with SAM
could form proper adhesion and fabricate MIP-QCM sensor
stably than MIP-QCM sensor without SAM, (c) bigger
weight of membrane improved sensor sensitivity, however
too big weight could not qualify itself as sensor, and (d)
optimization of material types and its composition was the
most significant issue for improvement of MIP-QCM
sensor. In fact, MIP-QCM sensor (BPA: 4-Vpy: EGDMA=
1: 4: 20, SAM=MUA) respond 20 Hz at BPA concentration
of 600 ppb.
Through these investigations, it is proved that each
optimal design will lead further improvement as sensor.
Acknowledgements
This work was partly supported by KEIO Life Con-
jugated Chemistry (LCC) Programme of the Centre of
Excellence (COE) Research Project in the 21st century
sponsored by Japanese Ministry of Education, Science,
Culture, and Sports.
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